Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from a peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Screening Of Food Dyes for Genotoxic Activity

Author:

R. B. Haveland-Smith And R. D. Combes

Year:

1980

Bibliographic source:

Food and Cosmetics Toxicology Volume 18, Issue 3, 1980, Pages 215-221

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

Modified liquid Fluctuation test

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate

EC Number:

224-924-0

EC Name:

Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate

Cas Number:

4553-89-3

Molecular formula:

C27H20N4O9S2.2Na

IUPAC Name:

disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate)
- Molecular formula: C27H20N4O9S2.2Na
- Molecular weight: 654.58 g/mol
- Substance type: Organic
- Physical state: Solid

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9

Test concentrations with justification for top dose:

5000 mg/L

Vehicle / solvent:

Deionised water

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: No data available
- Exposure duration: 72- 96 hrs
- Expression time (cells in growth medium): 72- 96 hrs
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: The food colour was made up in deionized water and membrane-sterilized prior to use. The test material was tested at its maximum sublethal concentration.

Evaluation criteria:

The test material was considered positive only if it resulted in significant more turbid tubes in a treated series when compared with an untreated set of tubes

Statistics:

Chi square test

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: allno mutagenic potential

Applicant's summary and conclusion

Conclusions:

Chocolate Brown HT ( Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate) is considered to be not genotoxic in the bacteria Salmonella typhimurium TA 1538 and Escherichia coli WP2 uvrA since no detectable effects were observed in the Modified liquid Fluctuation test.

Executive summary:

Bacterial gene mutation was studied using Modified liquid Fluctuation test. Chocolate Brown HT ( Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate) was used at a concentration of 5000 mg/kg and its genotoxic effect was studied in the bacteria Salmonella typhimurium TA 1538 and Escherichia coli WP2 uvrA. Te fluctuation assay in the liquid medium was conducted for 72-96 hrs at 25°C. The assay was performed for each bacterium in three separate experiments. The given test material is negative for inducing genotoxicity in the bacteria Salmonella typhimurium TA 1538 and Escherichia coli WP2 uvrA indicating that the substance is non-genotoxic.